基于CD19^(+)B细胞调控FAK/CAPN/PI3K通路研究新风胶囊改善类风湿关节炎机制

Mechanism of Xinfeng Capsules improving rheumatoid arthritis based on CD19^(+) B cells regulating FAK/CAPN/PI3K pathway

观察新风胶囊对类风湿关节炎(RA)B淋巴细胞、炎性介质、FAK/CAPN/PI3K通路的影响,探讨新风胶囊改善RA临床症状的机制。观察RA患者关节及全身症状,检测实验室指标[血红蛋白(HGB)、血小板计数(PLT)、血沉(ESR)、免疫球蛋白(Ig)G、IgA、IgM、类风湿因子(RF)、抗环瓜氨酸抗体(CCP-AB)、C反应蛋白(CRP)]。采用酶联免疫吸附法检测血清白细胞介素(IL)-1β、IL-10、IL-33、趋化因子5(CCL5)、血管内皮生长因子(VEGF),流式细胞术测定外周血CD3^(-)CD19^(+)B细胞,免疫印迹法检测FAK、p-FAK、CAPN、PI3K蛋白。结果显示,新风胶囊能明显改善RA关节及全身症状,提高临床疗效。且新风胶囊能升高HGB,降低PLT、CCP-AB、CRP、ESR指标,上调IL-10表达,下调IL-1β、IL-33、CCL5、VEGF、CD3^(-)CD19^(+)B细胞、FAK、p-FAK、CAPN、PI3K表达(P<0.01)。结果表明,新风胶囊可能通过下调CD3^(-)CD19^(+)B细胞表达,调控炎性细胞因子和趋化因子平衡,抑制FAK/CAPN/PI3K通路异常激活,改善RA临床症状。

To observe the effect of Xinfeng Capsules on rheumatoid arthritis(RA)B lymphocytes,inflammatory mediators,FAK/CAPN/PI3K pathway,in order to explore the mechanism of Xinfeng Capsules in improving clinical symptoms of RA.Joint and systemic symptoms of RA patients were observed,and laboratory indicators[hemoglobin(HGB),platelet count(PLT),erythrocyte sedimentation(ESR),immunoglobulin(Ig)G,IgA,IgM,rheumatoid factor(RF),anti-cyclic citrulline antibody(CCP-AB),C-reactive protein(CRP)]were detected.ELISA was used to detect serum interleukin(IL)-1β,IL-10,IL-33,chemokine 5(CCL5),and vascular endothelial growth factor(VEGF).CD3^(-)CD19^(+)B cells were measured by flow cytometry.Western blot was used to detect FAK,p-FAK,CAPN,PI3K protein.The results showed that Xinfeng Capsules could significantly alleviate RA joint and systemic symptoms and improve clinical efficacy.And Xinfeng Capsules could increase HGB,decrease PLT,CCP-AB,CRP,ESR index,upregulate IL-10 expression,and down-regulate IL-1β,IL-33,CCL5,VEGF,CD3^(-)CD19^(+)B cells,FAK,p-FAK,CAPN,PI3K expressions(P<0.01).Based on the above results,Xinfeng Capsules may reduce the expression of CD3^(-)CD19^(+),regulate the balance of inflammatory cytokines and chemokines,inhibit abnormal activation of FAK/CAPN/PI3K pathway,and improve clinical symptoms of RA.